Promotion of Oocyte Maturation and Embryonic Competence in Sheep by Viola alba subsp. sintenisii-Derived Bioactives

The impact of a hydrophenolic extract derived from Viola alba subsp. sintenisii (L.) . established on national Live collection of Viola- Zanjan, Iran (LCV) (36.6870267, 48.3881580) on the in vitro maturation (IVM) and fertilization of sheep cumulus-oocyte complexes (COCs) was investigated. Chemical analysis using LC-MS and GC-MS revealed the presence of biologically active compounds, including cyclotides, phenolic acids, and flavonoids, which have known roles in modulating cellular responses. COCs matured with the extract exhibited enhanced cumulus expansion, as evidenced by morphological observations and elevated expression of fshr (follicle-stimulating hormone receptor) and gdf9 (growth differentiation factor 9) genes. Specifically, the 50 µL concentration of the extract significantly upregulated these gene expressions, suggesting optimal follicular support and oocyte developmental competence. Furthermore, Rhodamine 123 staining showed that mitochondrial activity and distribution in oocytes significantly improved in the extract-treated groups, indicating enhanced energy metabolism and cytoplasmic maturation. Collectively, these findings highlight the efficacy of V. alba subsp. sintenisii extract VE in enhancing the IVM process, likely through the action of its bioactive compounds on key molecular and cellular pathways. These results provide a foundation for integrating natural compounds into assisted reproductive technologies to improve livestock production efficiency.